• Title/Summary/Keyword: Adiponectin gene

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Adiponectin Gene Cloning and Its Expression in Insect Cell Expression System

  • Yuh, In Suh;Sheffield, Lewis G.
    • Reproductive and Developmental Biology
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    • v.36 no.3
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    • pp.193-198
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    • 2012
  • This study was to examine expression of the recombinant full-length adiponectin (recombinant adiponectin) in insect ovarian cell culture system and to characterize structural properties of the recombinant adiponectin secreted in medium. Gene construct encoding the recombinant adiponectin contained N-terminal collagen-like domain (110 Amino Acids, AAs), C-terminal globular domain (137 AAs) and C-terminal peptides for detection with V5 antibody (26 AAs included adaptor peptide) and purification using the 6xHis tag (6 AAs). The approximate molecular weight of the product (monomer) was 35 kDa. Molecular mass species of the expressed recombinant adiponectin were monomer (~35 kDa), dimer (~70 kDa), trimer (~105 kDa) and hexamer (~210 kDa). The major secreted species were the LMW forms, such as monomer, dimer, and trimer. There was MMW of hexamer as minor form. HMW multimers (~300 kDa) were shown as a tracer or not detected on the SDS-PAGE in several experiments (data not shown). The multimer forms in this study were not compatible to those in animal or human serum and adipose tissue by other researcher's study in which the major multimer forms were HMW. By protein denaturing experiments with reducing reagent (${\beta}$-MeOH), anionic detergent (SDS) and heat ($95^{\circ}C$) on the SDS-PAGE, not all adiponectin multimers seemed to have disulfide bond linked structure to form multimers. The recombinant adiponectin which expressed in insect ovarian cell culture system seemed to have the limitation as full physiological regulator for the application to animal and human study.

Correlation of Gene Expression between Adiponectin and Glucose Transporter 4 in Mouse Adipose Tissue (생쥐 지방조직에서의 아디포넥틴과 포도당수송체-4 유전자 발현의 상관관계)

  • Lee, Yong-Ho
    • Journal of Life Science
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    • v.24 no.8
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    • pp.895-902
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    • 2014
  • Adiponectin has been known to improve insulin sensitivity and elicit glucose uptake via increased glucose transporter 4 (GLUT4) translocation. In the current study, mRNA expression levels of adiponectin and GLUT4 were measured in subcutaneous adipose tissue from C57BL/6 mice fed normal (ND) or high-fat diet (HFD) until 16, 26, 36, 47, or 77 weeks of age starting from 6 weeks of age. Expression levels were also measured in mice with calorie restriction (CR) and in thiazolidinedione (TZD) treated mice. Using quantitative real-time PCR, we demonstrated that GLUT4 expression in adipose tissue significantly decreased in HFD mice groups and increased in CR (p<0.05) and TZD (p=0.007) groups while there was no difference in adiponectin mRNA expression levels between experimental and control groups. General linear regression models were used to assess the association of gene expression levels between adiponectin and GLUT4 and to determine whether adiponectin affects GLUT4 transcription. mRNA expression levels of adiponectin and GLUT4 are significantly associated each other in mice fed a ND (p<0.0001) or HFD (p<0.0001), in groups separated into each age and diet, and CR group (p=0.002), but not in TZD group (p=0.73). These results demonstrated that gene expression of adiponectin and GLUT4 is strongly associated, suggesting that there is a common regulatory mechanism for adiponectin and GLUT4 gene expression and/or adiponectin has a direct role in GLUT4 gene expression in adipose tissue.

Association of the A-G Polymorphism in Porcine Adiponectin Gene with Fat Deposition and Carcass Traits

  • Dai, L.H.;Xiong, Y.Z.;Deng, C.Y.;Jiang, S.W.;Zuo, B.;Zheng, R.;Li, F.E.;Lei, M.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.6
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    • pp.779-783
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    • 2006
  • The adiponectin gene is known to be involved in the regulation of energy homeostasis involving food intake, carbohydrate and lipid metabolism. Human adiponectin gene polymorphisms have been recently reported to be associated with obesity, insulin sensitivity and the risk of type 2 diabetes. The present study was carried out to investigate the porcine adiponectin gene as a candidate gene for fat deposition and carcass traits. A mutation of A178G of the porcine adiponectin gene that resulted in substitution of the amino acid Isoleucine to Valine was identified. AcyI PCR-RFLP was used to detect the polymorphism of the genotypes in five different pig populations (Large White, Landrace, Duroc, Chinese breeds Meishan and Qingping). The A allele frequency was significantly higher among subjects from Chinsese lard type breeds, while the G allele was the only one present in those from Western lean type breeds. To determine if there was an association of the polymorphism with phenotypic variation, the mutation was tested in 267 pigs of the "Large $White{\times}Meishan$" F2 resource population. The results of association analyses showed significant associations of the genotypes with fat deposition and carcass traits. Allele G was significantly associated with increase in loin eye height, loin eye area and lean meat percentage and bone percentage, and decrease in fat mean percentage, ratio of lean to fat, shoulder fat thickness, 6-7 rib fat thickness, thorax-waist fat thickness and buttock fat thickness. The substitution of A178G (Ile60Val) happened to be located at amino acid 60 in the collagenous domain of porcine adiponectin which might affect the association into higher-order structures, and accordingly affect the posttranslational modifications and optimal biological activity of the multimeric forms. The identified functional polymorphism provides new evidence of adiponectin as an important candidate gene affecting fat deposition and carcass traits in pigs.

Adiponectin gene SNP 276G${\rightarrow}$T, nutrient intakes, and cardiovascular disease risk in Korean type 2 DM patients

  • Yu, So-Young;Ryu, Han-Kyoung;Park, Hee-Jung;Choi, Young-Ju;Huh, Kap-Bum;Kim, Wha-Young
    • Nutrition Research and Practice
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    • v.1 no.4
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    • pp.363-370
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    • 2007
  • Single nucleotide polymorphism (SNP) in adiponectin gene has been associated with insulin resistance, diabetes, and cardiovascular disease (CVD). This study was performed to investigate the association of SNP 276G${\rightarrow}$T at adiponectin gene with CVD risk factors in Korean type 2 diabetes mellitus (DM) patients. The subjects were 351 type 2 DM patients visited a DM clinic in Seoul, and the patients with known CVD were excluded. The adiponectin SNP 276G${\rightarrow}$T was analyzed and dietary intakes were assessed by a Food Frequency Questionnaire. The prevalence of G/G, G/T, and T/T genotype was 47.6%, 43.3%, and 9.1%, respectively. Male subjects with T/T genotype showed significantly lower level of adiponectin and HDL-cholesterol and significantly higher C-reactive protein (CRP) level compared to G/G and G/T genotypes. In G/G genotype, protein intake was negatively correlated to body weight, BMI, and waist circumference, and there were positive correlation between carbohydrate intake and BMI, waist-hip ratio, and ApoB/apoA-1 ratio in G/T genotype. However, in T/T genotype, there was no significant association between macronutrient intakes and anthropometric and hematological values. In conclusion, CVD risk would be high in type 2 DM patients with T/T genotype, and the association of macronutrient intakes with anthropometric and hematologic factors was different among the three adiponectin genotypes. These results may imply the need for different dietary management regime according to adiponectin genotype to lower CVD complications in Korean type 2 DM patients.

The Expression of Porcine Adiponectin and Stearoyl Coenzyme a Desaturase Genes in Differentiating Adipocytes

  • Wang, P.H.;Ko, Y.H.;Liu, B.H.;Peng, H.M.;Lee, M.Y.;Chen, C.Y.;Li, Y.C.;Ding, S.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.5
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    • pp.588-593
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    • 2004
  • The gene expression of porcine adiponectin and stearoyl coenzyme A desaturase (SCD) was investigated in this study. The partial gene sequences for adiponectin and SCD were amplified by RT-PCR from subcutaneous adipose tissue and cloned by TA cloning techniques. Sequences of these genes were determined and found to be highly homologous to that of other species, suggesting similar function of these genes as in other species. The transcripts of these adipocyte-related genes in pig tissues were measured by Northern analysis. The transcripts for adiponectin and SCD were highly expressed in porcine subcutaneous adipose tissue; the transcripts for SCD were also barely detected in the liver, but the greatest concentrations were in the adipose tissue. In porcine stromalvascular cells (S/V cells) cultured in vitro, transcripts for adiponectin and SCD increased gradually during adipocyte differentiation. The level of adipocyte adiponectin mRNA was associated with late adipocyte differentiation, indicating the gene may not be involved in adipocyte differentiation but has great importance in porcine adipocyte functions. The SCD transcripts were not detectable until 2 d after induction of adipocyte differentiation. It was highly expressed in differentiating porcine adipocytes (2 to 10 d after the induction of adipocyte differentiation), indicating a significant role of SCD in adipocytes.

Serum level of the adiponectin and adiponectin I164T polymorphism in hypertensive adolescents (고혈압 청소년에서의 혈청 adiponectin치와 adiponectin I164T 유전자다형성)

  • Lee, Jung Ah;Gil, Joo Hyun;Hong, Young Mi
    • Clinical and Experimental Pediatrics
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    • v.52 no.2
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    • pp.187-193
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    • 2009
  • Purpose : Adiponectin is a molecule that plays an important role in the metabolic syndrome. In addition, its concentration is known to be decreased in obesity, type 2 diabetes, and coronary artery disease. Although a relationship between hypertension and serum adiponectin concentrations has been reported by several authors, such findings continue to be debated. We investigated whether hypoadiponectinemia is related to hypertension in adolescents and studied the associated genetic polymorphism. Methods : Forty hypertensive adolescents (Age 16-17 years old) and twenty normotensive matched subjects were included. Serum adipo-nectin, insulin, renin, aldosterone and angiotensin converting enzyme (ACE) levels were compared. Their carotid intima-media thickness (cIMT) and pulse wave velocity (PWV) were measured. Polymorphisms of the adiponectin I164T gene were investigated using polymerase chain reaction (PCR). Results : The hypertensive adolescents had significantly greater cIMT and PWV. In addition, the serum aldosterone, renin and insulin levels were significantly higher in the hypertensive group. The plasma concentrations of adiponectin did not differ significantly between the two groups. TC genotype was not found in our study subjects; they all had the TT genotype of the adiponectine gene. Conclusion : The results of our study showed that adiponectin levels were not significantly different in adolescents with hypertension. There was no distinctive genetic polymorphism observed in this group of patients. Further large scale studies are needed to clarify the association between genetic variations and adiponectin in hypertensive adolescents.

Association of Adiponectin Polymorphisms with Type 2 Diabetes in Korean Population (한국인에서의 아디포넥틴의 유전자다형성과 제2형 당뇨병과의 연관성)

  • Yoo, Min;Kim, Su-Won
    • Journal of Life Science
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    • v.19 no.10
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    • pp.1495-1498
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    • 2009
  • Type 2-diabetes is a typical polygenic disease complex, for which several common risk alleles have been identified. Adiponectin, which modulates insulin resistance as well as glucose and lipid metabolism, has recently been associated with type 2-diabetes (T2DM). Therefore, we investigated the genotype for the T45G and G267T polymorphisms in adiponectin genes in the Korean population and compared genotypes of patients with those of a control group. 100 patients (63 male, 37 female), who previously underwent T2DM and 100 controls (36 male, 63 female) participated in this study. There was a strong association between T45G polymorphism in the adiponectin gene and T2DM. The present study shows that adiponectin T45G polymorphism may be associated with the pathogenesis of T2DM. Further studies with a larger population may be needed for the development of diagnostic methods at genetic levels such as DNA chip.

IL-2-enhanced NK Cell Cytotoxicity is Regulated by Adiponectin from Hypothalamo-pituitary-adrenal Axis (Adiponectin에 의한 IL-2 증가 자연살해세포 독성의 조절)

  • Kim, Keun-Young;Yang, Young
    • IMMUNE NETWORK
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    • v.6 no.1
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    • pp.6-12
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    • 2006
  • Background: The Hypothalamo-Pituitary-Adrenal (HPA) axis is an important regulator for the body's stress response. As a primary stress responsive system, HPA-axis secretes various neurotransmitters, hormones, and cytokines, which regulates the immune system. Natural killer (NK) cell which is plays an important role in the innate immune response, is specially decreased their numbers and loose cytolytic activity in response to stress. However, the effect of HPA-axis secreted proteins on NK cell activity has not been defined. Herein, we studied the effect of adrenal secreted adiponectin on NK cell cytotoxicity. Adiponectin which is well-known metabolic control protein, plays important roles in various diseases, including hypertension, cardiovascular diseases, inflammatory disorders, and cancer. Methods: Signal sequence trap was used to find stress novel secretory protein from HP A-axis. Selected adiponectin was treated mouse mature primary NK cells and then examined the effect of adiponectin to NK cell cytotoxicity and cytokine expression level. Results: We found that adiponectin which is secreted from adrenal gland, suppress IL-2 induced NK cell cytotoxicity. And also investigated cytolytic cytokines are suppressed by adiponectin. Conclusion: These data suggest that adiponectin inhibites NK cell cytotoxicity via suppression of cytotoxicity related target gene.

The Regulation of Chemerin and CMKLR1 Genes Expression by TNF-α, Adiponectin, and Chemerin Analog in Bovine Differentiated Adipocytes

  • Suzuki, Y.;Hong, Y.H.;Song, S.H.;Ardiyanti, A.;Kato, D.;So, K.H.;Katoh, K.;Roh, Sang-Gun
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.9
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    • pp.1316-1321
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    • 2012
  • Adipokines, adipocyte-derived protein, have important roles in various kinds of physiology including energy homeostasis. Chemerin, one of adipocyte-derived adipokines, is highly expressed in differentiated adipocytes and is known to induce macrophage chemotaxis and glucose intolerance. The objective of the present study was to investigate the changes of chemerin and the chemokine-like-receptor 1 (CMKLR1) gene expression levels during differentiation of the bovine adipocyte and in differentiated adipocytes treated with tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), adiponectin, leptin, and chemerin (peptide analog). The expression levels of the chemerin gene increased at d 6 and 12 of the differentiation period accompanied by increased cytoplasm lipid droplets. From d 6 onward, peroxisome proliferator-activated receptor-${\gamma}2$ (PPAR-${\gamma}2$) gene expression levels were significantly higher than that of d 0 and 3. In contrast, CMKLR1 expression levels decreased at the end of the differentiation period. In fully differentiated adipocytes (i.e. at d 12), the treatment of TNF-${\alpha}$ and adiponectin upregulated both chemerin and CMKLR1 gene expression levels, although leptin did not show such effects. Moreover, chemerin analog treatment was shown to upregulate chemerin gene expression levels regardless of doses. These results suggest that the expression of chemerin in bovine adipocyte might be regulated by chemerin itself and other adipokines, which indicates its possible role in modulating the adipokine secretions in adipose tissues.

ZFP36L1 and AUF1 Induction Contribute to the Suppression of Inflammatory Mediators Expression by Globular Adiponectin via Autophagy Induction in Macrophages

  • Shrestha, Aastha;Pun, Nirmala Tilija;Park, Pil-Hoon
    • Biomolecules & Therapeutics
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    • v.26 no.5
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    • pp.446-457
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    • 2018
  • Adiponectin, a hormone predominantly originated from adipose tissue, has exhibited potent anti-inflammatory properties. Accumulating evidence suggests that autophagy induction plays a crucial role in anti-inflammatory responses by adiponectin. However, underlying molecular mechanisms are still largely unknown. Association of Bcl-2 with Beclin-1, an autophagy activating protein, prevents autophagy induction. We have previously shown that adiponectin-induced autophagy activation is mediated through inhibition of interaction between Bcl-2 and Beclin-1. In the present study, we examined the molecular mechanisms by which adiponectin modulates association of Bcl-2 and Beclin-1 in macrophages. Herein, we demonstrated that globular adiponectin (gAcrp) induced increase in the expression of AUF1 and ZFP36L1, which act as mRNA destabilizing proteins, both in RAW 264.7 macrophages and primary peritoneal macrophages. In addition, gene silencing of AUF1 and ZFP36L1 caused restoration of decrease in Bcl-2 expression and Bcl-2 mRNA half-life by gAcrp, indicating crucial roles of AUF1 and ZFP36L1 induction in Bcl-2 mRNA destabilization by gAcrp. Moreover, knock-down of AUF1 and ZFP36L1 enhanced interaction of Bcl-2 with Beclin-1, and subsequently prevented gAcrp-induced autophagy activation, suggesting that AUF1 and ZFP36L1 induction mediates gAcrp-induced autophagy activation via Bcl-2 mRNA destabilization. Furthermore, suppressive effects of gAcrp on LPS-stimulated inflammatory mediators expression were prevented by gene silencing of AUF1 and ZFP36L1 in macrophages. Taken together, these results suggest that AUF1 and ZFP36L1 induction critically contributes to autophagy induction by gAcrp and are promising targets for anti-inflammatory responses by gAcrp.